【摘要】：目的:研究性激素合成通路中的關(guān)鍵酶之一的細(xì)胞色素P450c17a酶(CYP17A1)在三種人神經(jīng)膠質(zhì)瘤細(xì)胞系T98G、U87和U251中的表達(dá)水平,使用CYP17A1抑制劑Abiraterone干預(yù)膠質(zhì)瘤細(xì)胞株T98G,并探索其對膠質(zhì)瘤細(xì)胞增殖活性的影響,為涉及CYP17A1在膠質(zhì)瘤中的作用機(jī)制以及以CYP17A1為潛在靶點的腫瘤治療研究等方面奠定基礎(chǔ)。方法:(1)選取細(xì)胞活性良好的三種人腦膠質(zhì)瘤細(xì)胞系T98G、U87和U251為實驗樣本,采用Western blot和Real-time PCR的方法從蛋白質(zhì)水平和m RNA水平來檢測CYP17A1在三種膠質(zhì)瘤細(xì)胞系T98G、U87和U251的表達(dá)水平,并比較CYP17A1在三種膠質(zhì)瘤細(xì)胞系T98G、U87和U251間的蛋白質(zhì)和m RNA表達(dá)水平差異。(2)通過CYP17A1抑制劑Abiraterone干預(yù)膠質(zhì)瘤細(xì)胞株T98G,采用Real-time PCR,Western blot,cck8增殖實驗及流式細(xì)胞術(shù)實驗檢測Abiraterone干預(yù)對T98G中CYP17A1表達(dá)水平、細(xì)胞增殖活性與細(xì)胞凋亡活性的影響。結(jié)果:(1)Western blot結(jié)果顯示:檢測出CYP17A1在三種人腦膠質(zhì)瘤細(xì)胞系T98G、U251和U87中的相對表達(dá)量為(0.518±0.052)、(0.460±0.034)和(0.142±0.025)。人腦膠質(zhì)瘤細(xì)胞系T98G和U251中CYP17A1表達(dá)水平均明顯高于人腦膠質(zhì)瘤細(xì)胞系U87,差異有統(tǒng)計學(xué)意義(P0.05)。Real-time PCR檢測結(jié)果顯示:CYP17A1的m RNA在三種人腦膠質(zhì)瘤細(xì)胞系T98G、U251和U87中相對表達(dá)水平分別為(1.000±0.122)、(0.960±0.079)、(0.611±0.045)。其中,人腦膠質(zhì)瘤細(xì)胞系T98G中CYP17A1的表達(dá)水平高于人腦膠質(zhì)瘤細(xì)胞系U87,差異有統(tǒng)計學(xué)意義(P0.05);人腦膠質(zhì)瘤細(xì)胞系U251中CYP17A1的表達(dá)水平也要高于人腦膠質(zhì)瘤細(xì)胞系U87,差異同樣具有統(tǒng)計學(xué)意義(P0.05)。但是,Western blot和Real-time PCR都指出人腦膠質(zhì)瘤細(xì)胞系T98G和U251在CYP17A1在蛋白質(zhì)和m RNA的表達(dá)差異無統(tǒng)計學(xué)意義(P0.05);(2)Abiraterone對膠質(zhì)瘤的生物學(xué)活性產(chǎn)生抑制作用,能抑制膠質(zhì)瘤細(xì)胞的增殖,促進(jìn)膠質(zhì)瘤細(xì)胞的凋亡,并且這種抑制及促進(jìn)作用與Abiraterone的濃度相關(guān)(P0.05);(3)Abiraterone可以影響膠質(zhì)瘤細(xì)胞的CYP17A1表達(dá)水平,影響的效果也與Abiraterone濃度相關(guān)(P0.05)。結(jié)論:(1)神經(jīng)膠質(zhì)瘤細(xì)胞系T98G、U87和U251均表達(dá)CYP17A1,其中,膠質(zhì)瘤細(xì)胞系T98G和U251中CYP17A1的表達(dá)水平均明顯高于人腦膠質(zhì)瘤細(xì)胞系U87中CYP17A1的表達(dá)水平,但是在腦膠質(zhì)瘤細(xì)胞系T98G和U251中CYP17A1的表達(dá)水平并無明顯差異。(2)CYP17A1抑制劑能抑制膠質(zhì)瘤細(xì)胞的增殖,促進(jìn)膠質(zhì)瘤細(xì)胞的凋亡,并影響膠質(zhì)瘤細(xì)胞的CYP17A1表達(dá)水平。
[Abstract]:Aim: to study the expression of cytochrome P450c17a enzyme (CYP17A1), one of the key enzymes in sex hormone synthesis pathway, in three human glioma cell lines T98GnU87 and U251, and to use CYP17A1 inhibitor Abiraterone to interfere with the expression of cytochrome P450c17a enzyme (CYP17A1) in human glioma cell line T98G. The effects of CYP17A1 on the proliferation of glioma cells were explored, which laid a foundation for the mechanism of CYP17A1 in glioma and the study of tumor therapy with CYP17A1 as the potential target. Methods: (1) three kinds of glioma cell lines T98GU87 and U251 with good cell activity were selected as experimental samples. Western blot and Real-time PCR were used to detect CYP17A1 in three glioma cell lines T98G from protein level and m RNA level. The expression levels of U87 and U251 were compared between the three glioma cell lines T98GU87 and U251. (2) the CYP17A1 inhibitor Abiraterone was used to interfere with T98G and Real-time PCR,Western blot, was used to treat the glioma cell line T98G. Cck8 proliferation assay and flow cytometry were used to detect the effect of Abiraterone intervention on the expression of CYP17A1, cell proliferation activity and apoptosis activity in T98G. Results: (1) Western blot results showed that the relative expression of CYP17A1 was (0.518 鹵0.052), () 0.460 鹵0.034 and (0.142 鹵0.025) in three human glioma cell lines T98GnU251 and U87. The expression of CYP17A1 in human glioma cell line T98G and U251 was significantly higher than that in human glioma cell line U87 (P0.05). The results of Real-time PCR detection showed that the expression of m RNA of CYP17A1 in three kinds of human glioma cell lines T98G was significantly higher than that in human glioma cell line U87 (P0.05). The relative expression levels of U251 and U87 were (1.000 鹵0.122), (, 0.960 鹵0.079), (, 0.611 鹵0.045), respectively. The expression of CYP17A1 in human glioma cell line T98G was significantly higher than that in human glioma cell line U87 (P0.05). The expression of CYP17A1 in human glioma cell line U251 was also higher than that in human glioma cell line U87 (P0.05). However, both, Western blot and Real-time PCR indicated that there was no significant difference in the expression of T98G and U251 in CYP17A1 between protein and m RNA (P0.05). (2) Abiraterone inhibited the biological activity of glioma cells, inhibited the proliferation of glioma cells and promoted the apoptosis of glioma cells, and the inhibition and promotion effect was related to the concentration of Abiraterone (P0.05); (3) Abiraterone could affect the expression of CYP17A1 in glioma cells, and the effect was related to the concentration of Abiraterone (P0.05). Conclusion: (1) the expression of CYP17A1 in glioma cell line T98G and U251 was significantly higher than that in human glioma cell line U87 and U251, and the expression level of CYP17A1 in glioma cell line T98G and U251 was significantly higher than that in human glioma cell line U87. However, there was no significant difference in the expression of CYP17A1 between T98G and U251 glioma cells. (2) CYP17A1 inhibitor could inhibit the proliferation of glioma cells, promote the apoptosis of glioma cells, and affect the CYP17A1 expression level of glioma cells.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R739.41

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